COM-1 Promotes Homologous Recombination during Meiosis by Antagonizing Ku-Mediated Non-Homologous End Joining

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Successful completion of meiosis requires the induction and faithful repair of DNA double-strand breaks (DSBs). DSBs can be repaired via homologous recombination (HR) or non-homologous end joining (NHEJ), yet only repair via HR can generate the interhomolog crossovers (COs) needed for meiotic chromosome segregation. Here we identify COM-1, the homolog of CtIP/Sae2/Ctp1, as a crucial regulator of DSB repair pathway choice during Caenorhabditis elegans gametogenesis. COM-1–deficient germ cells repair meiotic DSBs via the error-prone pathway NHEJ, resulting in a lack of COs, extensive chromosomal aggregation, and near-complete embryonic lethality. In contrast to its yeast counterparts, COM-1 is not required for Spo11 removal and initiation of meiotic DSB repair, but instead promotes meiotic recombination by counteracting the NHEJ complex Ku. In fact, animals defective for both COM-1 and Ku are viable and proficient in CO formation. Further genetic dissection revealed that COM-1 acts parallel to the nuclease EXO-1 to promote interhomolog HR at early pachytene stage of meiotic prophase and thereby safeguards timely CO formation. Both of these nucleases, however, are dispensable for RAD-51 recruitment at late pachytene stage, when homolog-independent repair pathways predominate, suggesting further redundancy and/or temporal regulation of DNA end resection during meiotic prophase. Collectively, our results uncover the potentially lethal properties of NHEJ during meiosis and identify a critical role for COM-1 in NHEJ inhibition and CO assurance in germ cells.

Vyšlo v časopise: COM-1 Promotes Homologous Recombination during Meiosis by Antagonizing Ku-Mediated Non-Homologous End Joining. PLoS Genet 9(2): e32767. doi:10.1371/journal.pgen.1003276
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1003276

Souhrn

Zdroje

1. McKinnonPJ, CaldecottKW (2007) DNA strand break repair and human genetic disease. Annu Rev Genomics Hum Genet 8: 37–55.

2. LieberMR (2010) The mechanism of double-strand DNA break repair by the nonhomologous DNA end-joining pathway. Annu Rev Biochem 79: 181–211.

3. SymingtonLS, GautierJ (2011) Double-strand break end resection and repair pathway choice. Annu Rev Genet 45: 247–271.

4. ShaharOD, RamEV, ShimshoniE, HareliS, MeshorerE, et al. (2011) Live imaging of induced and controlled DNA double-strand break formation reveals extremely low repair by homologous recombination in human cells. Oncogene

5. KeeneyS, GirouxCN, KlecknerN (1997) Meiosis-specific DNA double-strand breaks are catalyzed by Spo11, a member of a widely conserved protein family. Cell 88: 375–384.

6. DernburgAF, McDonaldK, MoulderG, BarsteadR, DresserM, et al. (1998) Meiotic recombination in C. elegans initiates by a conserved mechanism and is dispensable for homologous chromosome synapsis. Cell 94: 387–398.

7. Hunter N (2006) Meiotic recombination; Aguilera A, and Rothstein, R., eds., editor. Heidelberg: Springer-Verlag.

8. MartinJS, WinkelmannN, PetalcorinMI, McIlwraithMJ, BoultonSJ (2005) RAD-51-dependent and -independent roles of a Caenorhabditis elegans BRCA2-related protein during DNA double-strand break repair. Mol Cell Biol 25: 3127–3139.

9. SmolikovS, EizingerA, HurlburtA, RogersE, VilleneuveAM, et al. (2007) Synapsis-defective mutants reveal a correlation between chromosome conformation and the mode of double-strand break repair during Caenorhabditis elegans meiosis. Genetics 176: 2027–2033.

10. AdamoA, CollisSJ, AdelmanCA, SilvaN, HorejsiZ, et al. (2010) Preventing nonhomologous end joining suppresses DNA repair defects of Fanconi anemia. Mol Cell 39: 25–35.

11. ClejanI, BoerckelJ, AhmedS (2006) Developmental modulation of nonhomologous end joining in Caenorhabditis elegans. Genetics 173: 1301–1317.

12. YunMH, HiomK (2009) CtIP-BRCA1 modulates the choice of DNA double-strand-break repair pathway throughout the cell cycle. Nature 459: 460–463.

13. ChenPL, LiuF, CaiS, LinX, LiA, et al. (2005) Inactivation of CtIP leads to early embryonic lethality mediated by G1 restraint and to tumorigenesis by haploid insufficiency. Mol Cell Biol 25: 3535–3542.

14. PenknerA, Portik-DobosZ, TangL, SchnabelR, NovatchkovaM, et al. (2007) A conserved function for a Caenorhabditis elegans Com1/Sae2/CtIP protein homolog in meiotic recombination. EMBO J 26: 5071–5082.

15. LimboO, ChahwanC, YamadaY, de BruinRA, WittenbergC, et al. (2007) Ctp1 is a cell-cycle-regulated protein that functions with Mre11 complex to control double-strand break repair by homologous recombination. Mol Cell 28: 134–146.

16. SartoriAA, LukasC, CoatesJ, MistrikM, FuS, et al. (2007) Human CtIP promotes DNA end resection. Nature 450: 509–514.

17. KeeneyS, KlecknerN (1995) Covalent protein-DNA complexes at the 5′ strand termini of meiosis-specific double-strand breaks in yeast. Proc Natl Acad Sci U S A 92: 11274–11278.

18. HartsuikerE, MizunoK, MolnarM, KohliJ, OhtaK, et al. (2009) Ctp1CtIP and Rad32Mre11 nuclease activity are required for Rec12Spo11 removal, but Rec12Spo11 removal is dispensable for other MRN-dependent meiotic functions. Mol Cell Biol 29: 1671–1681.

19. AdamoA, MontemauriP, SilvaN, WardJD, BoultonSJ, et al. (2008) BRC-1 acts in the inter-sister pathway of meiotic double-strand break repair. EMBO Rep 9: 287–292.

20. SeversonAF, LingL, van ZuylenV, MeyerBJ (2009) The axial element protein HTP-3 promotes cohesin loading and meiotic axis assembly in C. elegans to implement the meiotic program of chromosome segregation. Genes Dev 23: 1763–1778.

21. BhallaN, WynneDJ, JantschV, DernburgAF (2008) ZHP-3 acts at crossovers to couple meiotic recombination with synaptonemal complex disassembly and bivalent formation in C. elegans. PLoS Genet 4: e1000235 doi:10.1371/journal.pgen.1000235.

22. HillersKJ, VilleneuveAM (2003) Chromosome-wide control of meiotic crossing over in C. elegans. Curr Biol 13: 1641–1647.

23. YokooR, ZawadzkiKA, NabeshimaK, DrakeM, ArurS, et al. (2012) COSA-1 Reveals Robust Homeostasis and Separable Licensing and Reinforcement Steps Governing Meiotic Crossovers. Cell 149: 75–87.

24. YoudsJL, MetsDG, McIlwraithMJ, MartinJS, WardJD, et al. (2010) RTEL-1 enforces meiotic crossover interference and homeostasis. Science 327: 1254–1258.

25. AlpiA, PasierbekP, GartnerA, LoidlJ (2003) Genetic and cytological characterization of the recombination protein RAD-51 in Caenorhabditis elegans. Chromosoma 112: 6–16.

26. MetsDG, MeyerBJ (2009) Condensins regulate meiotic DNA break distribution, thus crossover frequency, by controlling chromosome structure. Cell 139: 73–86.

27. SaitoTT, YoudsJL, BoultonSJ, ColaiacovoMP (2009) Caenorhabditis elegans HIM-18/SLX-4 interacts with SLX-1 and XPF-1 and maintains genomic integrity in the germline by processing recombination intermediates. PLoS Genet 5: e1000735 doi:10.1371/journal.pgen.1000735.

28. MimitouEP, SymingtonLS (2010) Ku prevents Exo1 and Sgs1-dependent resection of DNA ends in the absence of a functional MRX complex or Sae2. EMBO J 29: 3358–3369.

29. SzankasiP, SmithGR (1995) A role for exonuclease I from S. pombe in mutation avoidance and mismatch correction. Science 267: 1166–1169.

30. ZakharyevichK, MaY, TangS, HwangPY, BoiteuxS, et al. (2010) Temporally and biochemically distinct activities of Exo1 during meiosis: double-strand break resection and resolution of double Holliday junctions. Mol Cell 40: 1001–1015.

31. HayashiM, ChinGM, VilleneuveAM (2007) C. elegans germ cells switch between distinct modes of double-strand break repair during meiotic prophase progression. PLoS Genet 3: e191 doi:10.1371/journal.pgen.0030191.

32. ColaiacovoMP, MacQueenAJ, Martinez-PerezE, McDonaldK, AdamoA, et al. (2003) Synaptonemal complex assembly in C. elegans is dispensable for loading strand-exchange proteins but critical for proper completion of recombination. Dev Cell 5: 463–474.

33. Martinez-PerezE, VilleneuveAM (2005) HTP-1-dependent constraints coordinate homolog pairing and synapsis and promote chiasma formation during C. elegans meiosis. Genes Dev 19: 2727–2743.

34. BickelJS, ChenL, HaywardJ, YeapSL, AlkersAE, et al. (2010) Structural maintenance of chromosomes (SMC) proteins promote homolog-independent recombination repair in meiosis crucial for germ cell genomic stability. PLoS Genet 6: e1001028 doi:10.1371/journal.pgen.1001028.

35. PasierbekP, JantschM, MelcherM, SchleifferA, SchweizerD, et al. (2001) A Caenorhabditis elegans cohesion protein with functions in meiotic chromosome pairing and disjunction. Genes Dev 15: 1349–1360.

36. ZouL, ElledgeSJ (2003) Sensing DNA damage through ATRIP recognition of RPA-ssDNA complexes. Science 300: 1542–1548.

37. BaillyA, GartnerA (2013) Germ cell apoptosis and DNA damage responses. Adv Exp Med Biol 757: 249–276.

38. Jaramillo-LambertA, EngebrechtJ (2010) A single unpaired and transcriptionally silenced X chromosome locally precludes checkpoint signaling in the Caenorhabditis elegans germ line. Genetics 184: 613–628.

39. ManfriniN, GueriniI, CitterioA, LucchiniG, LongheseMP (2010) Processing of meiotic DNA double strand breaks requires cyclin-dependent kinase and multiple nucleases. J Biol Chem 285: 11628–11637.

40. MoreauS, FergusonJR, SymingtonLS (1999) The nuclease activity of Mre11 is required for meiosis but not for mating type switching, end joining, or telomere maintenance. Mol Cell Biol 19: 556–566.

41. ChinGM, VilleneuveAM (2001) C. elegans mre-11 is required for meiotic recombination and DNA repair but is dispensable for the meiotic G(2) DNA damage checkpoint. Genes Dev 15: 522–534.

42. BaudrimontA, PenknerA, WoglarA, MamnunYM, HulekM, et al. (2011) A new thermosensitive smc-3 allele reveals involvement of cohesin in homologous recombination in C. elegans. PLoS ONE 6: e24799 doi:10.1371/journal.pone.0024799.

43. GarciaV, PhelpsSE, GrayS, NealeMJ (2011) Bidirectional resection of DNA double-strand breaks by Mre11 and Exo1. Nature 479: 241–244.

44. SunJ, LeeKJ, DavisAJ, ChenDJ (2011) Human Ku70/80 blocks Exonuclease1-mediated DNA resection in the presence of human Mre11 or Mre11/Rad50 complex. J Biol Chem

45. BlierPR, GriffithAJ, CraftJ, HardinJA (1993) Binding of Ku protein to DNA. Measurement of affinity for ends and demonstration of binding to nicks. J Biol Chem 268: 7594–7601.

46. FalzonM, FewellJW, KuffEL (1993) EBP-80, a transcription factor closely resembling the human autoantigen Ku, recognizes single- to double-strand transitions in DNA. J Biol Chem 268: 10546–10552.

47. ZhuangJ, JiangG, WillersH, XiaF (2009) Exonuclease function of human Mre11 promotes deletional nonhomologous end joining. J Biol Chem 284: 30565–30573.

48. LengsfeldBM, RattrayAJ, BhaskaraV, GhirlandoR, PaullTT (2007) Sae2 is an endonuclease that processes hairpin DNA cooperatively with the Mre11/Rad50/Xrs2 complex. Mol Cell 28: 638–651.

49. ShimEY, ChungWH, NicoletteML, ZhangY, DavisM, et al. (2010) Saccharomyces cerevisiae Mre11/Rad50/Xrs2 and Ku proteins regulate association of Exo1 and Dna2 with DNA breaks. EMBO J 29: 3370–3380.

50. KimHS, VijayakumarS, RegerM, HarrisonJC, HaberJE, et al. (2008) Functional interactions between Sae2 and the Mre11 complex. Genetics 178: 711–723.

51. RosuS, LibudaDE, VilleneuveAM (2011) Robust crossover assurance and regulated interhomolog access maintain meiotic crossover number. Science 334: 1286–1289.

52. RobertVJ, DavisMW, JorgensenEM, BessereauJL (2008) Gene conversion and end-joining-repair double-strand breaks in the Caenorhabditis elegans germline. Genetics 180: 673–679.

53. GoedeckeW, EijpeM, OffenbergHH, van AalderenM, HeytingC (1999) Mre11 and Ku70 interact in somatic cells, but are differentially expressed in early meiosis. Nat Genet 23: 194–198.

54. JiJ, TangD, WangK, WangM, CheL, et al. (2012) The role of OsCOM1 in homologous chromosome synapsis and recombination in rice meiosis. Plant J

55. KirkpatrickDT, FergusonJR, PetesTD, SymingtonLS (2000) Decreased meiotic intergenic recombination and increased meiosis I nondisjunction in exo1 mutants of Saccharomyces cerevisiae. Genetics 156: 1549–1557.

56. PetersonSE, LiY, ChaitBT, GottesmanME, BaerR, et al. (2011) Cdk1 uncouples CtIP-dependent resection and Rad51 filament formation during M-phase double-strand break repair. J Cell Biol 194: 705–720.

57. QvistP, HuertasP, JimenoS, NyegaardM, HassanMJ, et al. (2011) CtIP Mutations Cause Seckel and Jawad Syndromes. PLoS Genet 7: e1002310 doi:10.1371/journal.pgen.1002310.

58. BuntingSF, CallenE, WongN, ChenHT, PolatoF, et al. (2010) 53BP1 inhibits homologous recombination in Brca1-deficient cells by blocking resection of DNA breaks. Cell 141: 243–254.

59. KloostermanWP, GuryevV, van RoosmalenM, DuranKJ, de BruijnE, et al. (2011) Chromothripsis as a mechanism driving complex de novo structural rearrangements in the germline. Hum Mol Genet 20: 1916–1924.

60. ChiangC, JacobsenJC, ErnstC, HanscomC, HeilbutA, et al. (2012) Complex reorganization and predominant non-homologous repair following chromosomal breakage in karyotypically balanced germline rearrangements and transgenic integration. Nat Genet 44: 390–397.

61. BrennerS (1974) The genetics of Caenorhabditis elegans. Genetics 77: 71–94.

62. LeeMH, HollisSE, YooBH, NykampK (2011) Caenorhabditis elegans DNA-2 helicase/endonuclease plays a vital role in maintaining genome stability, morphogenesis, and life span. Biochem Biophys Res Commun 407: 495–500.